Classification of Tobaccos with Analytical Data of Nitrogen containing Compounds in Their Headspace Volatiles

In order to examine the relationship between tobacco aroma and the nitrogen-containing compounds in the tobacco headspace volatiles, the volatiles of flue-cured, Burley and Turkish tobaccos collected with Tenax GC were analyzed by a gas chromatograph equipped with a flame thermionic detector. Of the 21 compounds identified in the chromatogram, 2,5-dimethylpyridine, 2,4-dimethylpyridine, pyrrole and N-methylpyrrole have not been previously reported as present in the tobacco headspace volatiles. By principal component analysis and stepwise discriminant analysis, three principal components and discriminant functions with four peaks were obtained. The results of these statistical analyses classified the samples into each variety respectively.     

IL-17-mediated regulation of innate and acquired immune response against pulmonary Mycobacterium bovis bacille Calmette-Guerin infection

IL-17 is a cytokine that induces neutrophil-mediated inflammation, but its role in protective immunity against intracellular bacterial infection remains unclear. In the present study, we demonstrate that IL-17 is an important cytokine not only in the early neutrophil-mediated inflammatory response, but also in T cell-mediated IFN-gamma production and granuloma formation in response to pulmonary infection by Mycobacterium bovis bacille Calmette-Guérin (BCG). IL-17 expression in the BCG-infected lung was detected from the first day after infection and the expression depended on IL-23. Our observations indicated that gammadelta T cells are a primary source of IL-17. Lung-infiltrating T cells of IL-17-deficient mice produced less IFN-gamma in comparison to those from wild-type mice 4 wk after BCG infection. Impaired granuloma formation was also observed in the infected lungs of IL-17-deficient mice, which is consistent with the decreased delayed-type hypersensitivity response of the infected mice against mycobacterial Ag. These data suggest that IL-17 is an important cytokine in the induction of optimal Th1 response and protective immunity against mycobacterial infection.     

Antimicrobial susceptibility profile of Acinetobacter species isolated from blood cultures in two Japanese university hospitals

Carbapenem‐resistant Acinetobacter baumannii has rapidly spread worldwide. This study investigated antibiotic susceptibility and genotypic resistance of 123 consecutive blood culture isolates of Acinetobacter species collected between 2003 and 2011 in two Japanese hospitals. The isolates were assigned to 13 species. Carbapenem resistance was detected in four isolates. Only one A. baumannii isolate had bla_OXA‐23 together with ISAba1; the remaining three isolates had IMP‐1 metallo‐β‐lactamase. Quinolone resistance was detected in five isolates that had point mutations in the quinolone resistance‐determining region. The predominance of various non‐A. baumannii species and low prevalence of carbapenem resistance among blood culture isolates of Acinetobacter species in two Japanese hospitals were confirmed.     

NESH (Abi-3) is present in the Abi/WAVE complex but does not promote c-Abl-mediated phosphorylation

Abl interactor (Abi) was identified as an Abl tyrosine kinase-binding protein and subsequently shown to be a component of the macromolecular Abi/WAVE complex, which is a key regulator of Rac-dependent actin polymerization. Previous studies showed that Abi-1 promotes c-Abl-mediated phosphorylation of Mammalian Enabled (Mena) and WAVE2. In addition to Abi-1, mammals possess Abi-2 and NESH (Abi-3). In this study, we compared the three Abi proteins in terms of the promotion of c-Abl-mediated phosphorylation and the formation of Abi/WAVE complex. Although Abi-2, like Abi-1, promoted the c-Abl-mediated phosphorylation of Mena and WAVE2, NESH (Abi-3) had no such effect. This difference was likely due to their binding abilities as to c-Abl. Immunoprecipitation revealed that NESH (Abi-3) is present in the Abi/WAVE complex. Our results suggest that NESH (Abi-3), like Abi-1 and Abi-2, is a component of the Abi/WAVE complex, but likely plays a different role in the regulation of c-Abl.     

Radioimmunoassay for a novel lignan-related hypocholesterolemic agent, S-8921, in human plasma after high-performance liquid chromatography purification and in human urine after immunoaffinity extraction

The novel compound methyl-1-(3,4-dimethoxyphenyl)-3-(3-ethylvaleryl)-4-hydroxy-6,7,8-trimethoxy-2-naphthoate (S-8921) has hypocholesterolemic activity in animals and is expected to exhibit a similar activity in human. Reversed-phase high-performance liquid chromatography (HPLC) separation followed by radioimmunoassay (RIA) for human plasma samples (HPLC–RIA) and immunoaffinity extraction (IAE) followed by RIA for human urine samples (IAE–RIA) were developed for investigation of S-8921 behavior in clinical studies. For the RIA, antisera from rabbit and a radioiodine-labelled S-8921 were prepared by immunizing a conjugate of S-8921 with bovine serum albumin and by the Bolton and Hunter method, respectively. HPLC–RIA using a semi-micro column was very sensitive, that is a 0.05 ng/ml limit of quantitation in human plasma, and specific for unchanged form of S-8921. IAE–RIA using a centrifugal filtration tube completely eliminated the matrix effect of human urine, and was very feasible. The limit of quantitation was 0.10 ng/ml. RIA detection following HPLC or IAE proved to be very useful for the pharmaceutical analysis of extremely low drug concentrations in body fluids.     

Presence of antibiotic-producing streptococci in squid sauce during shiokara fermentation

The bacteriology of fresh and boiled squid sauce containing muscle has been studied. Bacterial numbers greatly increased only in the latter. Streptococcus spp. isolated from the fresh sauce showed anti-microbial activities to nine authentic strains tested, but not to Bacillus spp. which predominate in the fermented sauce.     

Ectopic synaptic ribbons in dendrites of mouse retinal ON- and OFF-bipolar cells

The ectopic distribution of synaptic ribbons in dendrites of mouse retinal bipolar cells was examined by using genetic ablation of metabotropic glutamate receptor subtype 6 (mGluR6), electron microscopy, and immunocytochemistry. Ectopic ribbons were observed in dendrites of rod and ON-cone bipolar cells in the mGluR6-deficient mouse but not in those of wild-type mice. The number of rod spherules facing the ectopic ribbons in mGluR6-deficient rod bipolar dendrites increased gradually during early growth and reached a plateau level of about 20% at 12 weeks. These ectopic ribbons were immunopositive for RIBEYE, a ribbon-specific protein, but the associated vesicles were immunonegative for synaptophysin, a synaptic-vesicle-specific protein. The presence of ectopic ribbons was correlated with an increase in the roundness of the invaginating dendrites of the rod bipolar cells. We further confirmed ectopic ribbons in dendrites of OFF-cone bipolar cells in wild-type retinas. Of the four types of OFF-cone bipolar cells (T1–T4), only the T2-type, which had a greater number of synaptic ribbons at the axon terminal and a thicker axon cylinder than the other types, had ectopic ribbons. Light-adapted experiments revealed that, in wild-type mice under enhanced-light adaptation (considered similar to the mGluR6-deficient state), the roundness in the invaginating dendrites and axon terminals of rod bipolar cells increased, but no ectopic ribbons were detected. Based on these findings and known mechanisms for neurotransmitter release and protein trafficking, the possible mechanisms underlying the ectopic ribbons are discussed on the basis of intracellular transport for the replenishment of synaptic proteins.     

Effects of fatty acids on superoxide radical generation in leukocytes

Acetylated ferricytochrome c was employed for the detection of superoxide radicals (O₂^?) generated both in intact cells and in subcellular fractions of leukocytes. Certain saturated fatty acids, myristate in particular, induced the production of O₂^? in leukocytes, suggesting a correlation between the formation of O₂^? and the hydrophobic interaction of fatty acids with the leukocyte plasma membrane. As compared with O₂^? radical generation from phagocytizing leukocytes, a greater stimulation of O_2? formation was observed in cells in which myristate was added. The enhanced activity which generated O₂^? in the cell-free system was located in a particulate fraction but not in the cytosol. The rate of O₂^? generation in the particulate fraction was higher in the presence of NADPH than in the presence of NADH. The effects of reagents such as KCN, 2,4-dichlorophenol and aminotriazole on the O₂^? generation in this fraction are examined and the nature of the O₂^? generating system is discussed.     

Carbohydrate structures of a normal counterpart of the carcinoembryonic antigen produced by colon epithelial cells of normal adults

Normal Faecal antigen-2 (NFA-2) and non-specific crossreactingantigen-2 (NCA-2), cross-reacting with anticarcinoembryonicantigen (CEA) antibodies, were found in normal human faececand meconium, respectively. Because NFA-2, NCA-2 and CEA areconsidered as the same gene products, NFA-2 and NCA-2 shouldbe normal counterparts of CEA produced by colon epithelial cellsof normal adults and fetuses, respectively. Comparison of sugarchain structures of these three antigens is indispensable inorder to unravel the stnaural alteration induced by malignanttransformation and development of colon epithelial cells. Thesugar chain structures of CEA (Yamashita,K. et al., Cancer Res.,47,3451–3459,1987) and NCA-2 (Yarnashita,K. et al., J.Bid Chem, 264,17873-17881,1989) were previously reported. Inthis paper, the structures of the oligosaccharides releasedfrom four NFA-2 samples by hydrazinolysis were studied by meansof lectin-affinity column chromatography, endo- and em-glycosidasedigestion, methylation analysis, hydrazinolys-nitrous acid deaminationand electrospray ionization mass spectrometry. NFA-2 contains24–27 mol of N-linked sugar chains/molecule, which issimilar to NCA-2 (27 mol) and CEA (24–27 mol). In contrastto CEA, which contains {small tilde} 8% high-mannose-type sugarchains all sugar chains of NFA-2 are mono- to tetra-antennarycomplex-type chains having four types of tri-mannosyl cores,with or without bisecting N-acetylglucosamine and fucose residues.The structures of their outer chain moieties comprise Galß1